title: Production and characterisation of a monoclonal antibody that recognises the chicken CSF1 receptor and confirms that expression is restricted to macrophage-lineage cells.
abstract: Macrophages contribute to innate and acquired immunity as well as many aspects of homeostasis and development. Studies of macrophage biology and function in birds have been hampered by a lack of definitive cell surface markers. As in mammals, avian macrophages proliferate and differentiate in response to CSF1 and IL34, acting through the shared receptor, CSF1R. CSF1R mRNA expression in the chicken is restricted to macrophages and their progenitors. To expedite studies of avian macrophage biology, we produced an avian CSF1R-Fc chimeric protein and generated a monoclonal antibody (designated ROS-AV170) against the chicken CSF1R using the chimeric protein as immunogen. Specific binding of ROS-AV170 to CSF1R was confirmed by FACS, ELISA and immunohistochemistry on tissue sections. CSF1 down-regulated cell surface expression of the CSF1R detected with ROS-AV170, but the antibody did not block CSF1 signalling. Expression of CSF1R was detected on the surface of bone marrow progenitors only after culture in the absence of CSF1, and was induced during macrophage differentiation. Constitutive surface expression of CSF1R distinguished monocytes from other myeloid cells, including heterophils and thrombocytes. This antibody will therefore be of considerable utility for the study of chicken macrophage biology.
authors.affiliation: The Roslin Institute & Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
authors.forename: Carla
authors.initials: C
authors.lastname: Garcia-Morales
authors.forename: Lisa
authors.initials: L
authors.lastname: Rothwell
authors.forename: Lindsey
authors.initials: L
authors.lastname: Moffat
authors.forename: Valerie
authors.initials: V
authors.lastname: Garceau
authors.forename: Adam
authors.initials: A
authors.lastname: Balic
authors.forename: Helen M
authors.initials: HM
authors.lastname: Sang
authors.forename: Pete
authors.initials: P
authors.lastname: Kaiser
authors.forename: David A
authors.initials: DA
authors.lastname: Hume
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001854:Bone Marrow Cells / Q000276:immunology; D016466:CHO Cells; D002454:Cell Differentiation / Q000276:immunology; D002460:Cell Line; D019070:Cell Lineage / Q000276:immunology; D002645:Chickens / Q000276:immunology*; D003412:Cricetulus; D005260:Female; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009000:Monocytes / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D016173:Macrophage Colony-Stimulating Factor; D016186:Receptor, Macrophage Colony-Stimulating Factor
keywords: Antibody; BMDM; CSF1R; FACS; Macrophages; PBMC; WB; Western blot; bone marrow-derived macrophages; colony stimulating factor 1 receptor; fluorescence-activated cell sorting; mAb; monoclonal antibody; peripheral blood monocytes